PROJECT SUMMARY/ABSTRACT This proposal describes a training plan to develop my academic and scientific career focusing on the experi- mental evolution of pancreatic cancer. The award's purpose is to advance my scientific independence with a research program seeking to quantify multiple facets of pancreatic cancer evolution. During the K99 award pe- riod, my mentor will be Dr. Iacobuzio-Donahue, a world-expert in pancreatic cancer genetics, evolution and pathology. Dr. Iacobuzio-Donahue has a well-established interest in cancer genetics and experimental strate- gies to define evolutionary patterns. My co-mentors will be Drs. Sohrab Shah and Dana Pe'er. Each has a long track record of mentorship ? many of the scientists and clinicians they have mentored have advanced to fruitful academic careers. In addition, Memorial Sloan Kettering (MSK) will provide support and resources for conduct- ing the research outlined in this proposal, including my training plan and successful transition to academic in- dependence. The research strategy of this proposal seeks to 1) enhance my scientific knowledge and technical skills related to tumor evolution, and 2) develop a novel approach to isolate and quantify multiple facets of pancreatic can- cer. This work will also extend my scientific background from the phylogenetics of human tumors to experi- mental strategies and novel evolutionary patterns of cancer evolution. Specific Aim 1 seeks to quantify tem- poral dynamics of nutrient deprived pancreatic metastatic cells with real-time sampling and single cell sequenc- ing, and Specific Aim 2 will define spatial diversity and migration of pancreatic cancer subclones using autopsy tissues, patient derived xenografts, single cell sequencing and phylogenetic analysis. During the two-year K99 period of this award experiments and sample collection for Specific Aims 1 and 2 will be completed. In the 3- year R00 award period, validation experiments and sequencing analysis will be subsequently completed to de- termine the evolutionary dynamics of pancreatic cancer cells. The long-term objective of this proposed research is to better define various influences of pancreatic cancer evolution. The proposed experimental strategy has the potential to 1) quantify tumor evolution one aspect at a time and 2) replay evolutionary events to delineate reproducibility. Given the aggressiveness of pancreatic cancer, new strategies like the proposed approach are urgently needed to quantify and thwart the evolutionary influences driving this disease, ultimately to improve patient treatment and outcomes.